April 27, 2000 

VETERINARY SERVICES MEMORANDUM NO. 800.70 

Subject: Rabies Vaccine Immunogenicity Test Protocols 

To: Biologics Licensees, Permittees, and Applicants 

Directors, Center for Veterinary Biologics 

I. PURPOSE 

This memorandum provides guidelines for preparing Rabies Vaccine immunogenicity 

test protocols to be filed with the Animal and Plant Health Inspection Service (APHIS) 

according to 9 CFR 113.209(b) and 113.312(b). 

II. CANCELLATION 

This memorandum cancels Veterinary Services Memorandum No. 800.70 dated May 29, 

1985. 

III. BACKGROUND 

According to the standard test requirements for Rabies Vaccine, Killed Virus, in 9 CFR 

113.209(b) and for Rabies Vaccine, Live Virus, in 113.312(b), the immunogenicity of the 

vaccine must be established in each species for which the vaccine is recommended and 

the tests must be conducted in accordance with a protocol filed with APHIS before 

initiation of the tests. The following guidelines are provided to assist applicants in 

developing protocols that will be acceptable to APHIS. 

IV. GUIDELINES 

A. General Information

1. Outline of Production - Submit an Outline of Production for the test 

product as specified in 9 CFR 114.9. 

2. Test Facilities - Provide the location of animal holding and challenge 

test facilities. Describe these facilities and provide authorization for access for 

CVB inspections if not on licensed premises. 

3. Test Dates - Provide tentative dates for onset, challenge, and 

termination of each test. 

4. CVB Observation - Submit protocols to Center for Veterinary 

Biologics-Licensing and Policy Development at least 60 days before the firm 
Veterinary Services Memorandum No. 800.70 2 

intends to initiate the study to provide adequate time for CVB to make 

arrangements for CVB personnel to observe the administration of the product 

including conduct of vaccine dilutions, if applicable. CVB personnel observing 

the study may select and authenticate samples of the vaccine, diluent, and diluted 

vaccine for use in conducting confirmatory tests at Center for Veterinary 

Biologics-Laboratory. CVB, if not observing the study, may request the 

submission of vaccine, diluent, and diluted vaccine for use in conducting these 

tests. 

B. Detailed Test Procedures

Provide detailed test procedures that describe the following: 

1. Vaccine Preparation - Indicate how the vaccine is prepared for use in 

the trial, including all details regarding dilution, if conducted. 

2. Method(s) of administration - Include the route and method of 

vaccination. 

3. Test animals - The Center for Veterinary Biologics prefers that animals 

less than 1 year of age and of random breed and sources be used. Indicate how 

contemporary controls are held throughout the test period. 

4. Collection of Serum Samples - Indicate the provisions made for 

collection of serum samples from test animals throughout the study. 

5. Identity of Challenge Virus - Identify the challenge virus to be used. If 

from a source other than CVB-L, provide a complete description. 

6. The Challenge Method - Include the mouse LD5O of the challenge 

material and the intramuscular site to be used for challenge. 

7. Tests Used to Determine Cause of Death - Describe the tests that will 

be used to determine the cause of death of any test animal. 

C. How to File

Detailed protocols and general information should be sent to: 

Center for Veterinary Biologics 

Licensing and Policy Development 

510 South 17th Street, Suite 104 

Ames, IA 50010-8197 
Veterinary Services Memorandum No. 800.70 3 

/s/ 

Alfonso Torres 

Deputy Administrator 

Veterinary Services